Basic research on anti-tumor effects of Interferon-α on hepatocellular carcinoma

干扰素-α抗肝癌作用的基础研究

• 批准号: 13670233
• 负责人: YANO Hirohisa
• 金额: $ 1.92万
• 依托单位: KURUME UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670233/
• 关键词: Hepatocellular carcinoma; Interferon-α; Apoptosis; Caspase; Cell line; Nude mouse; 5-Fu; Interferon-α subtypes; インターフェロン-α; コンセンサスインターフェロン; アポトーシス; ヌードマウス; 血管新生; 抗癌剤; 肝癌; 細胞株; カスペース(caspase); cytochrome c; RT-PCR; Western blotting; インタ

[Purpose] In this study, we examined that the effects of IFN-α subtypes on the growth of human liver cancer cell lines in vitro ; growth inhibitory effects of IFN-α and 5-Fu in vitro ; mechanisms of apoptosis induction by IFN-α; and the effects of IFN-α alone or IFN-α plus 5-Fu on the growth of human liver cancer cell lines transplanted In nude mice.[Results] 1. When we examined the effects of five kinds of recombinant IFN-α subtypes (α1, α2, α5, α8, α10; concentrations: 0-1024 IU/mL; 24-96 hours) on the growth of 13 human liver cancer cell lines, the strongest anti-proliferative effects was noted In IFN-α5 subtype, and followed by α8, α10, α2, α1. 2. No synergistic growth Inhibitory effects could be recognized when IFN-α and 5-Fu were simultaneously added t o the culture of liver cancer cells. 3. Natural human IFN-α induced activation of caspase-3, -7, -8, and -9, and proteolysis of PARP in IFN-α-induced apoptosis-sensitive cell lines at various degrees. In addition, release of cytochrome C and Smac/Diablo from mitochondria into cytoplasm was identified in IFN-α-treated cells time-dependently. These findings suggest mitochondria apoptosis induction pathway is involved in the IFN-α-induced apoptosis. 4. Subcutaneous IFN-αCon1 injection (0.01, 0.1, 1μg/mouse/day; 0.01 μg/mouse = clinical dosesage) for consecutive 14 days dose-and time-dependently suppressed the HCC tumor growth in nude mice. Finally, the tumor volumes of mice received 0.01, 0.1, and 1 μg/mouse of IFN-αCon1 decreased to about 60%, 25%,and 0-10%, respectively, of control tumor volumes. The number of apoptotic cells significantly Increased and the number of blood vessels significantly decreased with the increase of IFN-αCon1 dose. 5. Synergistic effect of IFN-α plus 5-Fu was identified In nude mouse tumors. [Conclusions] These data Indicate the potential clinical application of certain types of IFN-α in the prevention and treatment of HCC.

【目的】本研究探讨IFN-α亚型对人肝癌细胞系体外生长的影响；IFN-α和5-Fu的体外生长抑制作用；IFN-α诱导细胞凋亡的机制；以及IFN-α单独或IFN-α + 5-Fu对裸鼠移植人肝癌细胞株生长的影响。[结果]1。我们检测了5种重组IFN-α亚型（α1、α2、α5、α8、α10，浓度0 ~ 1024 IU/mL， 24 ~ 96 h）对13株人肝癌细胞株生长的影响，其中IFN-α5亚型的抗增殖作用最强，其次是α8、α10、α2、α1。2. 当IFN-α和5-Fu同时加入肝癌细胞培养时，未发现协同生长抑制作用。3. 在IFN-α诱导的凋亡敏感细胞系中，天然人IFN-α不同程度地诱导caspase-3、-7、-8和-9的激活以及PARP的蛋白水解。此外，在IFN-α-处理的细胞中，细胞色素C和Smac/Diablo从线粒体释放到细胞质中具有时间依赖性。提示线粒体凋亡诱导途径参与了IFN-α-诱导的细胞凋亡。4. 皮下注射IFN-αCon1（0.01、0.1、1μg/小鼠/天，0.01 μg/小鼠为临床剂量）连续14 d抑制裸鼠肝癌肿瘤生长。结果表明，注射0.01、0.1和1 μg/只IFN-αCon1后，小鼠肿瘤体积分别减少至对照肿瘤体积的60%、25%和0-10%左右。随着IFN-αCon1剂量的增加，凋亡细胞数量显著增加，血管数量显著减少。5. 在裸鼠肿瘤中发现IFN-α + 5-Fu的协同作用。[结论]这些数据提示某些类型的IFN-α在预防和治疗HCC中的潜在临床应用。